Triazole-containing macrocyclic HCV serine protease inhibitors

ABSTRACT

The present invention discloses compounds of formula I, II and III or pharmaceutically acceptable salts, esters, or prodrugs thereof: 
                         
which inhibit serine protease activity, particularly the activity of hepatitis C virus (HCV) NS3-NS4A protease. Consequently, the compounds of the present invention interfere with the life cycle of the hepatitis C virus and are also useful as antiviral agents. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HCV infection. The invention also relates to methods of treating an HCV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

RELATED APPLICATIONS

This application claims the benefit of U.S. provisional application No.61/013,724 filed on Dec. 14, 2007. The contents of the above applicationare incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to novel hepatitis C virus (HCV) proteaseinhibitor compounds having antiviral activity against HCV and useful inthe treatment of HCV infections. More particularly, the inventionrelates to triazole-containing macrocyclic compounds, compositionscontaining such compounds and methods for using the same, as well asprocesses for making such compounds.

BACKGROUND OF THE INVENTION

HCV is the principal cause of non-A, non-B hepatitis and is anincreasingly severe public health problem both in the developed anddeveloping world. It is estimated that the virus infects over 200million people worldwide, surpassing the number of individuals infectedwith the human immunodeficiency virus (HIV) by nearly five fold. HCVinfected patients, due to the high percentage of individuals inflictedwith chronic infections, are at an elevated risk of developing cirrhosisof the liver, subsequent hepatocellular carcinoma and terminal liverdisease. HCV is the most prevalent cause of hepatocellular cancer andcause of patients requiring liver transplantations in the western world.

There are considerable barriers to the development of anti-HCVtherapeutics, which include, but are not limited to, the persistence ofthe virus, the genetic diversity of the virus during replication in thehost, the high incident rate of the virus developing drug-resistantmutants, and the lack of reproducible infectious culture systems andsmall-animal models for HCV replication and pathogenesis. In a majorityof cases, given the mild course of the infection and the complex biologyof the liver, careful consideration must be given to antiviral drugs,which are likely to have significant side effects.

Only two approved therapies for HCV infection are currently available.The original treatment regimen generally involves a 3-12 month course ofintravenous interferon-α (IFN-α), while a new approved second-generationtreatment involves co-treatment with IFN-α and the general antiviralnucleoside mimics like ribavirin. Both of these treatments suffer frominterferon related side effects as well as low efficacy against HCVinfections. There exists a need for the development of effectiveantiviral agents for treatment of HCV infection due to the poortolerability and disappointing efficacy of existing therapies.

In a patient population where the majority of individuals arechronically infected and asymptomatic and the prognoses are unknown, aneffective drug would desirably possess significantly fewer side effectsthan the currently available treatments. The hepatitis C non-structuralprotein-3 (NS3) is a proteolytic enzyme required for processing of theviral polyprotein and consequently viral replication. Despite the hugenumber of viral variants associated with HCV infection, the active siteof the NS3 protease remains highly conserved thus making its inhibitionan attractive mode of intervention. Recent success in the treatment ofHIV with protease inhibitors supports the concept that the inhibition ofNS3 is a key target in the battle against HCV.

HCV is a flaviridae type RNA virus. The HCV genome is enveloped andcontains a single strand RNA molecule composed of circa 9600 base pairs.It encodes a polypeptide comprised of approximately 3010 amino acids.

The HCV polyprotein is processed by viral and host peptidase into 10discreet peptides which serve a variety of functions. There are threestructural proteins, C, E1 and E2. The P7 protein is of unknown functionand is comprised of a highly variable sequence. There are sixnon-structural proteins. NS2 is a zinc-dependent metalloproteinase thatfunctions in conjunction with a portion of the NS3 protein. NS3incorporates two catalytic functions (separate from its association withNS2): a serine protease at the N-terminal end, which requires NS4A as acofactor, and an ATP-ase-dependent helicase function at the carboxylterminus. NS4A is a tightly associated but non-covalent cofactor of theserine protease.

The NS3-NS4A protease is responsible for cleaving four sites on theviral polyprotein. The NS3-NS4A cleavage is autocatalytic, occurring incis. The remaining three hydrolyses, NS4A-NS4B, NS4B-NS5A and NS5A-NS5Ball occur in trans. NS3 is a serine protease which is structurallyclassified as a chymotrypsin-like protease. While the NS serine proteasepossesses proteolytic activity by itself, the HCV protease enzyme is notan efficient enzyme in terms of catalyzing polyprotein cleavage. It hasbeen shown that a central hydrophobic region of the NS4A protein isrequired for this enhancement. The complex formation of the NS3 proteinwith NS4A seems necessary to the processing events, enhancing theproteolytic efficacy at all of the sites.

A general strategy for the development of antiviral agents is toinactivate virally encoded enzymes, including NS3, that are essentialfor the replication of the virus. Current efforts directed toward thediscovery of NS3 protease inhibitors were reviewed by S. Tan, A. Pause,Y. Shi, N. Sonenberg, Hepatitis C Therapeutics: Current Status andEmerging Strategies, Nature Rev. Drug Discov. 1, 867-881 (2002).

SUMMARY OF THE INVENTION

The present invention relates to triazole-containing macrocycliccompounds and pharmaceutically acceptable salts, esters or prodrugsthereof, and methods of using the same to treat hepatitis C infection ina subject in need of such therapy. Macrocyclic compounds of the presentinvention interfere with the life cycle of the hepatitis C virus and arealso useful as antiviral agents. The present invention further relatesto pharmaceutical compositions comprising the aforementioned compounds,salts, esters or prodrugs for administration to a subject suffering fromHCV infection. The present invention further features pharmaceuticalcompositions comprising a compound of the present invention (or apharmaceutically acceptable salt, ester or prodrug thereof) and anotheranti-HCV agent, such as interferon (e.g., alpha-interferon,beta-interferon, consensus interferon, pegylated interferon, or albuminor other conjugated interferon), ribavirin, amantadine, another HCVprotease inhibitor, or an HCV polymerase, helicase or internal ribosomeentry site inhibitor. The invention also relates to methods of treatingan HCV infection in a subject by administering to the subject apharmaceutical composition of the present invention. The presentinvention further relates to pharmaceutical compositions comprising thecompounds of the present invention, or pharmaceutically acceptablesalts, esters, or prodrugs thereof, in combination with apharmaceutically acceptable carrier or excipient.

In one embodiment of the present invention there are disclosed compoundsrepresented by Formulas I, II or III, or pharmaceutically acceptablesalts, esters, or prodrugs thereof:

Wherein

A is absent or selected from (C═O), S(O)₂, C(═N—OR₁) or C(═N—CN);wherein R₁ is selected at each occurrence from the group consisting of:

(i) hydrogen;

(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(iii) heterocycloalkyl or substituted heterocycloalkyl;

(iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eacheach containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;—C₃-C₁₂ cyclically, or substituted —C₃-C₁₂ cyclically; —C₃-C₁₂cycloalkenyl, or substituted —C₃-C₁₂ cycloalkenyl;

L₂₀₁ is absent or selected from C₁-C₈ alkylene, C₂-C₈ alkenylene, orC₂-C₈ alkynylene containing 0, 1, 2, or 3 heteroatoms selected from O,S, or N; substituted C₁-C₈ alkylene, substituted C₂-C₈ alkenylene, orsubstituted C₂-C₈ alkynylene each containing 0, 1, 2, or 3 heteroatomsselected from O, S or N; C₃-C₁₂ cycloalkylene, or substituted C₃-C₁₂cycloalkylene each containing 0, 1, 2, or 3 heteroatoms selected from O,S or N; C₃-C₁₂ cycloalkenylene, or substituted C₃-C₁₂ cycloalkenyleneeach containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;

M is absent or selected from O, S or NR₁;

L₁₀₁ is absent or selected from C₁-C₈ alkylene, C₂-C₈ alkenylene, orC₂-C₈ alkynylene each containing 0, 1, 2, or 3 heteroatoms selected fromO, S, or N; substituted C₁-C₈ alkylene, substituted C₂-C₈ alkenylene, orsubstituted C₂-C₈ alkynylene each containing 0, 1, 2, or 3 heteroatomsselected from O, S or N; C₃-C₁₂ cycloalkylene, or substituted C₃-C₁₂cycloalkylene each containing 0, 1, 2, or 3 heteroatoms selected from O,S or N; C₃-C₁₂ cycloalkenylene, or substituted C₃-C₁₂ cycloalkenyleneeach containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;

Z₁₀₁ is absent or selected from aryl, substituted aryl, heteroaryl, orsubstituted heteroaryl;

W₁₀₁ is absent, or selected from C₁-C₈ alkylene, C₂-C₈ alkenylene, orC₂-C₈ alkynylene, C(O)NR₁, C(O), aryl, substituted aryl, heteroaryl, orsubstituted heteroaryl;

W₂₀₁ is selected from hydrogen, halogen, C₁-C₆ alkyl, C₃-C₁₂ cyclically,aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,substituted heteroaryl, heteroarylalkyl, substituted heteroarylalkyl,heterocycloalkyl, or substituted heterocycloalkyl;

alternatively, W₁₀₁ and W₂₀₁ taken together with the carbon atoms towhich they are attached to form a cyclic moiety selected from aryl,substituted aryl, heteroaryl, or substituted heteroaryl;

R and R′ are independently selected from the group consisting of:

(i) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cyclically, or substituted —C₃-C₁₂ cyclically; —C₄-C₁₂ alkylcycloalkyl,or substituted —C₄-C₁₂ alkylcycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; —C₄-C₁₂ alkylcycloalkenyl, orsubstituted —C₄-C₁₂ alkylcycloalkenyl;

(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(iii) heterocycloalkyl or substituted heterocycloalkyl;

(iv) hydrogen; deuterium;

G is selected from —OH, —NR₃R₄, —NHS(O)₂—R₂, —NH(SO₂)NR₃R₄;

R₂ is selected from:

(i) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(ii) heterocycloalkyl or substituted heterocycloalkyl;

(iii) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S or N, substituted —C₁-C₈alkyl, substituted -C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cyclically, or substituted —C₃-C₁₂ cyclically; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl;

R₃ and R₄ are independently selected from:

(i) hydrogen;

(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(iii) heterocycloalkyl or substituted heterocycloalkyl;

(iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cyclically, or substituted —C₃-C₁₂ cyclically; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl;

m=0, 1, or 2; and

m′=1 or 2.

In another embodiment, the present invention features pharmaceuticalcompositions comprising a compound of the invention, or apharmaceutically acceptable salt, ester or prodrug thereof. In stillanother embodiment of the present invention there are disclosedpharmaceutical compositions comprising a therapeutically effectiveamount of a compound of the invention, or a pharmaceutically acceptablesalt, ester or prodrug thereof, in combination with a pharmaceuticallyacceptable carrier or excipient. In yet another embodiment of theinvention are methods of treating a hepatitis C infection in a subjectin need of such treatment with said pharmaceutical compositions.

DETAILED DESCRIPTION OF THE INVENTION

A first embodiment of the invention is a compound represented by FormulaI or Formula II or Formula III as described above, or a pharmaceuticallyacceptable salt, ester or prodrug thereof, alone or in combination witha pharmaceutically acceptable carrier or excipient.

Other embodiments of the invention are compounds represented by FormulaIV, V or VI:

-   -   or a pharmaceutically acceptable salt, ester or prodrug thereof,        alone or in combination with a pharmaceutically acceptable        carrier or excipient, where R, R′, A, L₂₀₁, M, L₁₀₁, Z₁₀₁, W₁₀₁,        W₂₀₁ and G are as defined in the previous embodiment.

Representative compounds of the invention include, but are not limitedto, the following compounds (Table 1) according to Formula VII whereinR, L-W, Z, R′ and G are delineated for each example in Table 1.

TABLE 1 (VII)

Example # R L-W Z R′ G 1.

2.

3.

4.

OH 5.

6.

7.

8.

OH 9.

10.

11.

12.

OH 13.

14.

15.

16.

OH 17.

18.

19.

20.

OH 21.

22.

23.

24.

OH 25.

26.

27.

28.

OH 29.

30.

31.

32.

OH 33.

34.

35.

36.

OH 37.

38.

39.

40.

OH 41.

42.

43.

44.

OH 45.

46.

47.

48.

OH 49.

50.

51.

52.

OH 53.

54.

55.

56.

OH 57.

58.

59.

60.

OH 61.

62.

63.

64.

OH 65.

66.

67.

68.

OH 69.

70.

71.

72.

OH 73.

74.

75.

76.

OH 77.

78.

79.

80.

OH 81.

82.

83.

84.

OH 85.

86.

87.

88.

OH 89.

90.

91.

92.

OH 93.

94.

95.

96.

OH 97.

98.

99.

100.

OH 101.

102.

103.

104.

OH 105.

106.

107.

108.

OH 109.

110.

111.

112.

OH 113.

114.

115.

116.

OH 117.

118.

119.

120.

OH 121.

122.

123.

124.

OH 125.

126.

127.

128.

OH 129.

130.

131.

132.

OH 133.

134.

135.

136.

OH 137.

138.

139.

140.

OH 141.

142.

143.

144.

OH

The present invention also features pharmaceutical compositionscomprising a compound of the present invention, or a pharmaceuticallyacceptable salt, ester or prodrug thereof.

Compounds of the present invention can be administered as the soleactive pharmaceutical agent, or used in combination with one or moreagents to treat or prevent hepatitis C infections or the symptomsassociated with HCV infection. Other agents to be administered incombination with a compound or combination of compounds of the inventioninclude therapies for disease caused by HCV infection that suppressesHCV viral replication by direct or indirect mechanisms. These includeagents such as host immune modulators (for example, interferon-alpha,pegylated interferon-alpha, interferon-beta, interferon-gamma, CpGoligonucleotides and the like), or antiviral compounds that inhibit hostcellular functions such as inosine monophosphate dehydrogenase (forexample, ribavirin and the like). Also included are cytokines thatmodulate immune function. Also included are vaccines comprising HCVantigens or antigen adjuvant combinations directed against HCV. Alsoincluded are agents that interact with host cellular components to blockviral protein synthesis by inhibiting the internal ribosome entry site(IRES) initiated translation step of HCV viral replication or to blockviral particle maturation and release with agents targeted toward theviroporin family of membrane proteins such as, for example, HCV P7 andthe like. Other agents to be administered in combination with a compoundof the present invention include any agent or combination of agents thatinhibit the replication of HCV by targeting proteins of the viral genomeinvolved in the viral replication. These agents include but are notlimited to other inhibitors of HCV RNA dependent RNA polymerase such as,for example, nucleoside type polymerase inhibitors described in WO0190121(A2), or U.S. Pat. No. 6,348,587B1 or WO0160315 or WO0132153 ornon-nucleoside inhibitors such as, for example, benzimidazole polymeraseinhibitors described in EP 1162196A1 or WO0204425 or inhibitors of HCVprotease such as, for example, peptidomimetic type inhibitors such asBILN2061 and the like or inhibitors of HCV helicase.

Other agents to be administered in combination with a compound of thepresent invention include any agent or combination of agents thatinhibit the replication of other viruses for co-infected individuals.These agent include but are not limited to therapies for disease causedby hepatitis B (HBV) infection such as, for example, adefovir,lamivudine, and tenofovir or therapies for disease caused by humanimmunodeficiency virus (HIV) infection such as, for example, proteaseinhibitors: ritonavir, lopinavir, indinavir, nelfinavir, saquinavir,amprenavir, atazanavir, tipranavir, TMC-114, fosamprenavir; reversetranscriptase inhibitors: zidovudine, lamivudine, didanosine, stavudine,tenofovir, zalcitabine, abacavir, efavirenz, nevirapine, delavirdine,TMC-125; integrase inhibitors: L-870812, S-1360, or entry inhibitors:enfuvirtide (T-20), T-1249.

Accordingly, one aspect of the invention is directed to a method fortreating or preventing an infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents selected from the group consisting of a host immunemodulator and a second antiviral agent, or a combination thereof, with atherapeutically effective amount of a compound or combination ofcompounds of the invention, or a pharmaceutically acceptable salt,stereoisomer, tautomer, prodrug, salt of a prodrug, or combinationthereof. Examples of the host immune modulator are, but not limited to,interferon-alpha, pegylated-interferon-alpha, interferon-beta,interferon-gamma, a cytokine, a vaccine, and a vaccine comprising anantigen and an adjuvant, and said second antiviral agent inhibitsreplication of HCV either by inhibiting host cellular functionsassociated with viral replication or by targeting proteins of the viralgenome.

Further aspect of the invention is directed to a method of treating orpreventing infection caused by an RNA-containing virus comprisingco-administering to a patient in need of such treatment an agent orcombination of agents that treat or alleviate symptoms of HCV infectionincluding cirrhosis and inflammation of the liver, with atherapeutically effective amount of a compound or combination ofcompounds of the invention, or a pharmaceutically acceptable salt,stereoisomer, tautomer, prodrug, salt of a prodrug, or combinationthereof. Yet another aspect of the invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents that treat patients for disease caused by hepatitis B(HBV) infection, with a therapeutically effective amount of a compoundor a combination of compounds of the invention, or a pharmaceuticallyacceptable salt, stereoisomer, tautomer, prodrug, salt of a prodrug, orcombination thereof. An agent that treats patients for disease caused byhepatitis B (HBV) infection may be for example, but not limited thereto,L-deoxythymidine, adefovir, lamivudine or tenfovir, or any combinationthereof. Example of the RNA-containing virus includes, but not limitedto, hepatitis C virus (HCV).

Another aspect of the invention provides a method of treating orpreventing infection caused by an RNA-containing virus comprisingco-administering to a patient in need of such treatment one or moreagents that treat patients for disease caused by human immunodeficiencyvirus (HIV) infection, with a therapeutically effective amount of acompound or a combination of compounds of the invention, or apharmaceutically acceptable salt, stereoisomer, tautomer, prodrug, saltof a prodrug, or combination thereof. The agent that treats patients fordisease caused by human immunodeficiency virus (HIV) infection mayinclude, but is not limited thereto, ritonavir, lopinavir, indinavir,nelfmavir, saquinavir, amprenavir, atazanavir, tipranavir, TMC-114,fosamprenavir, zidovudine, lamivudine, didanosine, stavudine, tenofovir,zalcitabine, abacavir, efavirenz, nevirapine, delavirdine, TMC-125,L-870812, S-1360, enfuvirtide (T-20) or T-1249, or any combinationthereof. Example of the RNA-containing virus includes, but not limitedto, hepatitis C virus (HCV). In addition, the present invention providesthe use of a compound or a combination of compounds of the invention, ora therapeutically acceptable salt form, stereoisomer, or tautomer,prodrug, salt of a prodrug, or combination thereof, and one or moreagents selected from the group consisting of a host immune modulator anda second antiviral agent, or a combination thereof, to prepare amedicament for the treatment of an infection caused by an RNA-containingvirus in a patient, particularly hepatitis C virus. Examples of the hostimmune modulator are, but not limited to, interferon-alpha, pegylated-interferon-alpha, interferon-beta, interferon-gamma, a cytokine, avaccine, and a vaccine comprising an antigen and an adjuvant, and saidsecond antiviral agent inhibits replication of HCV either by inhibitinghost cellular functions associated with viral replication or bytargeting proteins of the viral genome.

When used in the above or other treatments, combination of compound orcompounds of the invention, together with one or more agents as definedherein above, can be employed in pure form or, where such forms exist,in pharmaceutically acceptable salt form, prodrug, salt of a prodrug, orcombination thereof. Alternatively, such combination of therapeuticagents can be administered as a pharmaceutical composition containing atherapeutically effective amount of the compound or combination ofcompounds of interest, or their pharmaceutically acceptable salt form,prodrugs, or salts of the prodrug, in combination with one or moreagents as defined hereinabove, and a pharmaceutically acceptablecarrier. Such pharmaceutical compositions can be used for inhibiting thereplication of an RNA-containing virus, particularly Hepatitis C virus(HCV), by contacting said virus with said pharmaceutical composition. Inaddition, such compositions are useful for the treatment or preventionof an infection caused by an RNA-containing virus, particularlyHepatitis C virus (HCV).

Hence, further aspect of the invention is directed to a method oftreating or preventing infection caused by an RNA-containing virus,particularly a hepatitis C virus (HCV), comprising administering to apatient in need of such treatment a pharmaceutical compositioncomprising a compound or combination of compounds of the invention or apharmaceutically acceptable salt, stereoisomer, or tautomer, prodrug,salt of a prodrug, or combination thereof, one or more agents as definedhereinabove, and a pharmaceutically acceptable carrier.

When administered as a combination, the therapeutic agents can beformulated as separate compositions which are given at the same time orwithin a predetermined period of time, or the therapeutic agents can begiven as a single unit dosage form.

Antiviral agents contemplated for use in such combination therapyinclude agents (compounds or biologicals) that are effective to inhibitthe formation and/or replication of a virus in a mammal, including butnot limited to agents that interfere with either host or viralmechanisms necessary for the formation and/or replication of a virus ina mammal. Such agents can be selected from another anti-HCV agent; anHIV inhibitor; an HAV inhibitor; and an HBV inhibitor.

Other anti-HCV agents include those agents that are effective fordiminishing or preventing the progression of hepatitis C relatedsymptoms or disease. Such agents include but are not limited toimmunomodulatory agents, inhibitors of HCV NS3 protease, otherinhibitors of HCV polymerase, inhibitors of another target in the HCVlife cycle and other anti-HCV agents, including but not limited toribavirin, amantadine, levovirin and viramidine.

Immunomodulatory agents include those agents (compounds or biologicals)that are effective to enhance or potentiate the immune system responsein a mammal. Immunomodulatory agents include, but are not limited to,inosine monophosphate dehydrogenase inhibitors such as VX-497(merimepodib, Vertex Pharmaceuticals), class I interferons, class IIinterferons, consensus interferons, asialo-interferons pegylatedinterferons and conjugated interferons, including but not limited tointerferons conjugated with other proteins including but not limited tohuman albumin. Class I interferons are a group of interferons that allbind to receptor type I, including both naturally and syntheticallyproduced class I interferons, while class II interferons all bind toreceptor type II. Examples of class I interferons include, but are notlimited to, [alpha]-, [beta]-, [delta]-, [omega]-, and[tau]-interferons, while examples of class II interferons include, butare not limited to, [gamma]-interferons.

Inhibitors of HCV NS3 protease include agents (compounds or biologicals)that are effective to inhibit the function of HCV NS3 protease in amammal. Inhibitors of HCV NS3 protease include, but are not limited to,those compounds described in WO 99/07733, WO 99/07734, WO 00/09558, WO00/09543, WO 00/59929, WO 03/064416, WO 03/064455, WO 03/064456, WO2004/030670, WO 2004/037855, WO 2004/039833, WO 2004/101602, WO2004/101605, WO 2004/103996, WO 2005/028501, WO 2005/070955, WO2006/000085, WO 2006/007700 and WO 2006/007708 (all by BoehringerIngelheim), WO 02/060926, WO 03/053349, W003/099274, WO 03/099316, WO2004/032827, WO 2004/043339, WO 2004/094452, WO 2005/046712, WO2005/051410, WO 2005/054430 (all by BMS), WO 2004/072243, WO2004/093798, WO 2004/113365, WO 2005/010029 (all by Enanta), WO2005/037214 (Intermune) and WO 2005/051980 (Schering), and thecandidates identified as VX-950, ITMN-191 and SCH 503034.

Inhibitors of HCV polymerase include agents (compounds or biologicals)that are effective to inhibit the function of an HCV polymerase. Suchinhibitors include, but are not limited to, non-nucleoside andnucleoside inhibitors of HCV NS5B polymerase. Examples of inhibitors ofHCV polymerase include but are not limited to those compounds describedin: WO 02/04425, WO 03/007945, WO 03/010140, WO 03/010141, WO2004/064925, WO 2004/065367, WO 2005/080388 and WO 2006/007693 (all byBoehringer Ingelheim), WO 2005/049622 (Japan Tobacco), WO 2005/014543(Japan Tobacco),WO 2005/012288 (Genelabs), WO 2004/087714 (IRBM), WO03/101993 (Neogenesis), WO 03/026587 (BMS), WO 03/000254 (JapanTobacco), and WO 01/47883 (Japan Tobacco), and the clinical candidatesXTL-2125, HCV 796, R-1626 and NM 283.

Inhibitors of another target in the HCV life cycle include agents(compounds or biologicals) that are effective to inhibit the formationand/or replication of HCV other than by inhibiting the function of theHCV NS3 protease. Such agents may interfere with either host or HCVviral mechanisms necessary for the formation and/or replication of HCV.Inhibitors of another target in the HCV life cycle include, but are notlimited to, entry inhibitors, agents that inhibit a target selected froma helicase, a NS2/3 protease and an internal ribosome entry site (IRES)and agents that interfere with the function of other viral targetsincluding but not limited to an NS5A protein and an NS4B protein.

It can occur that a patient may be co-infected with hepatitis C virusand one or more other viruses, including but not limited to humanimmunodeficiency virus (HIV), hepatitis A virus (HAV) and hepatitis Bvirus (HBV). Thus also contemplated is combination therapy to treat suchco-infections by co-administering a compound according to the presentinvention with at least one of an HIV inhibitor, an HAV inhibitor and anHBV inhibitor.

According to yet another embodiment, the pharmaceutical compositions ofthe present invention may further comprise inhibitor(s) of other targetsin the HCV life cycle, including, but not limited to, helicase,polymerase, metalloprotease, and internal ribosome entry site (IRES).

According to another embodiment, the pharmaceutical compositions of thepresent invention may further comprise another anti-viral,anti-bacterial, anti-fungal or anti-cancer agent, or an immunemodulator, or another thearapeutic agent.

According to still another embodiment, the present invention includesmethods of treating viral infection such as, but not limited to,hepatitis C infections in a subject in need of such treatment byadministering to said subject an effective amount of a compound of thepresent invention or a pharmaceutically acceptable salt, ester, orprodrug thereof.

According to a further embodiment, the present invention includesmethods of treating hepatitis C infections in a subject in need of suchtreatment by administering to said subject an anti-HCV virally effectiveamount or an inhibitory amount of a pharmaceutical composition of thepresent invention.

An additional embodiment of the present invention includes methods oftreating biological samples by contacting the biological samples withthe compounds of the present invention.

Yet a further aspect of the present invention is a process of making anyof the compounds delineated herein employing any of the synthetic meansdelineated herein.

DEFINITIONS

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “viral infection” refers to the introduction of a virus intocells or tissues, e.g., hepatitis C virus (HCV). In general, theintroduction of a virus is also associated with replication. Viralinfection may be determined by measuring virus antibody titer in samplesof a biological fluid, such as blood, using, e.g., enzyme immunoassay.Other suitable diagnostic methods include molecular based techniques,such as RT-PCR, direct hybrid capture assay, nucleic acid sequence basedamplification, and the like. A virus may infect an organ, e.g., liver,and cause disease, e.g., hepatitis, cirrhosis, chronic liver disease andhepatocellular carcinoma.

The term “anti-cancer agent” refers to a compound or drug capable ofpreventing or inhibiting the advancement of cancer. Examples of suchagents include cis-platin, actinomycin D, doxorubicin, vincristine,vinblastine, etoposide, amsacrine, mitoxantrone, tenipaside, taxol,colchicine, cyclosporin A, phenothiazines or thioxantheres.

The term “anti-fungal agent” shall used to describe a compound which maybe used to treat a fungus infection other than 3-AP, 3-AMP or prodrugsof 3-AP and 3-AMP according to the present invention. Anti-fungal agentsaccording to the present invention include, for example, terbinafine,fluconazole, itraconazole, posaconazole, clotrimazole, griseofulvin,nystatin, tolnaftate, caspofungin, amphotericin B, liposomalamphotericin B, and amphotericin B lipid complex.

The term “antibacterial agent” refers to both naturally occurringantibiotics produced by microorganisms to suppress the growth of othermicroorganisms, and agents synthesized or modified in the laboratorywhich have either bactericidal or bacteriostatic activity, e.g.,β-lactam antibacterial agents, glycopeptides, macrolides, quinolones,tetracyclines, and aminoglycosides. In general, if an antibacterialagent is bacteriostatic, it means that the agent essentially stopsbacterial cell growth (but does not kill the bacteria); if the agent isbacteriocidal, it means that the agent kills the bacterial cells (andmay stop growth before killing the bacteria).

The term “immune modulator” refers to any substance meant to alter theworking of the humoral or cellular immune system of a subject. Suchimmune modulators include inhibitors of mast cell-mediated inflammation,interferons, interleukins, prostaglandins, steroids, cortico-steroids,colony-stimulating factors, chemotactic factors, etc.

The term “C₁-C₆ alkyl,” or “C₁-C₈ alkyl,” as used herein, refer tosaturated, straight- or branched-chain hydrocarbon radicals containingbetween one and six, or one and eight carbon atoms, respectively.Examples of C₁-C₆ alkyl radicals include, but are not limited to,methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl,n-hexyl radicals; and examples of C₁-C₈ alkyl radicals include, but arenot limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl,neopentyl, n-hexyl, heptyl, octyl radicals.

The term “C₂-C₆ alkenyl,” or “C₂-C₈ alkenyl,” as used herein, denote amonovalent group derived from a hydrocarbon moiety by the removal of asingle hydrogen atom wherein the hydrocarbon moiety has at least onecarbon-carbon double bond and contains from two to six, or two to eightcarbon atoms, respectively. Alkenyl groups include, but are not limitedto, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,heptenyl, octenyl and the like.

The term “C₂-C₆ alkynyl,” or “C₂-C₈ alkynyl,” as used herein, denote amonovalent group derived from a hydrocarbon moiety by the removal of asingle hydrogen atom wherein the hydrocarbon moiety has at least onecarbon-carbon triple bond and contains from two to six, or two to eightcarbon atoms, respectively. Representative alkynyl groups include, butare not limited to, for example, ethynyl, 1-propynyl, 1-butynyl,heptynyl, octynyl and the like.

The term “C₃-C₈-cycloalkyl”, or “C₃-C₁₂-cycloalkyl,” as used herein,denotes a monovalent group derived from a monocyclic or polycyclicsaturated carbocyclic ring compound by the removal of a single hydrogenatom where the saturated carbocyclic ring compound has from 3 ot 8, orfrom 3 to 12, ring atoms, respectively. Examples of C₃-C₈-cyclicallyinclude, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cyclopentyl and cyclooctyl; and examples ofC₃-C₁₂-cyclically include, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2]octyl.

The term “C₃-C₈-cycloalkenyl”, or “C₃-C₁₂-cycloalkenyl” as used herein,denote a monovalent group derived from a monocyclic or polycycliccarbocyclic ring compound having at least one carbon-carbon double bondby the removal of a single hydrogen atom where the carbocyclic ringcompound has from 3 ot 8, or from 3 to 12, ring atoms, respectively.Examples of C₃-C₈-cycloalkenyl include, but not limited to,cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl,cyclooctenyl, and the like; and examples of C₃-C₁₂-cycloalkenyl include,but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like.

The term “aryl,” as used herein, refers to a mono- or bicycliccarbocyclic ring system having one or two aromatic rings including, butnot limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, idenyland the like.

The term “arylalkyl,” as used herein, refers to a C₁-C₃ alkyl or C₁-C₆alkyl residue attached to an aryl ring. Examples include, but are notlimited to, benzyl, phenethyl and the like.

The term “heteroaryl,” as used herein, refers to a mono-, bi-, ortri-cyclic aromatic radical or ring having from five to ten ring atomsof which at least one ring atom is selected from S, O and N; wherein anyN or S contained within the ring may be optionally oxidized. Heteroarylincludes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl,pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl,thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and thelike.

The term “heteroarylalkyl,” as used herein, refers to a C₁-C₃ alkyl orC₁-C₆ alkyl residue residue attached to a heteroaryl ring. Examplesinclude, but are not limited to, pyridinylmethyl, pyrimidinylethyl andthe like.

The term “substituted” as used herein, refers to independent replacementof one, two, or three or more of the hydrogen atoms thereon withsubstituents including, but not limited to, —F, —Cl, —Br, —I, —OH,protected hydroxy, —NO₂, —CN, —NH₂, N₃, protected amino, alkoxy,thioalkoxy, oxo, -halo-C₁-C12-alkyl, -halo-C₂-C₁₂-alkenyl,-halo-C₂-C₁₂-alkynyl, -halo-C₃-C₁₂-cyclically, —NH—C₁-C₁₂-alkyl,—NH—C₂-C₁₂-alkenyl, —NH—C₂-C₁₂-alkynyl, —NH—C₃-C₁₂-cyclically, —NH-aryl,—NH-heteroaryl, —NH-heterocycloalkyl, -dialkylamino, -diarylamino,-diheteroarylamino, —O—C₁-C₁₂-alkyl, —O—C₂-C₁₂-alkenyl,—O—C₂-C₁₂-alkynyl, —O—C₃-C₁₂-cyclically, —O-aryl, —O-heteroaryl,—O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₁₂-alkenyl,—C(O)—C₂-C₁₂-alkynyl, —C(O)—C₃-C₁₂-cyclically, —C(O)-aryl,—C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl,—CONH—C₂-C₁₂-alkenyl, —CONH—C₂-C₁₂-alkynyl, —CONH—C₃-C₁₂-cyclically,—CONH-aryl, —CONH-heteroaryl, —CONH-heterocycloalkyl,—OCO₂—C₁-C₁₂-alkyl, —OCO₂-C₂-C₁₂-alkenyl, —OCO₂—C₂-C₁₂-alkynyl,—OCO₂—C₃-C₁₂-cyclically, —OCO₂-aryl, —OCO₂-heteroaryl,—OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₁₂-alkenyl, —OCONH—C₂-C₁₂-alkynyl, —OCONH—C₃-C₁₂-cyclically,—OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₁₂-alkenyl, —NHC(O)—C₂-C₁₂-alkynyl,—NHC(O)—C₃-C₁₂-cyclically, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₁₂-alkenyl,—NHCO₂—C₂-C₁₂-alkynyl, —NHCO₂—C₃-C₁₂-cyclically, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₁₂-alkenyl,—NHC(O)NH—C₂-C₁₂-alkynyl, —NHC(O)NH—C₃-C₁₂-cyclically, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₁₂-alkenyl,—NHC(S)NH—C₂-C₁₂-alkynyl, —NHC(S)NH—C₃-C₁₂-cyclically, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₁₂-alkenyl,—NHC(NH)NH—C₂-C₁₂-alkynyl, —NHC(NH)NH—C₃-C₁₂-cyclically,—NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₁₂-alkenyl, —NHC(NH)—C₂-C₁₂-alkynyl,—NHC(NH)—C₃-C₁₂-cyclically, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₁₂-alkenyl, —C(NH)NH—C₂-C₁₂-alkynyl,—C(NH)NH—C₃-C₁₂-cyclically, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₁₂-alkenyl,—S(O)—C₂-C₁₂-alkynyl, —S(O)—C₃-C₁₂-cyclically, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₂-C₁₂-alkynyl, —SO₂NH—C₃-C₁₂-cyclically,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₁₂-alkenyl, —NHSO₂—C₂-C₁₂-alkynyl,—NHSO₂—C₃-C₁₂-cyclically, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cyclically,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₁₂-alkenyl, —S—C₂-C₁₂-alkynyl,—S—C₃-C₁₂-cyclically, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl,methylthiomethyl, or -L′-R′, wherein L′ is C₁-C₆alkylene,C₂-C₆alkenylene or C₂-C₆alkynylene, and R′ is aryl, heteroaryl,heterocyclic, C₃-C₁₂cyclically or C₃-C₁₂cycloalkenyl. It is understoodthat the aryls, heteroaryls, alkyls, and the like can be furthersubstituted. In some cases, each substituent in a substituted moiety isadditionally optionally substituted with one or more groups, each groupbeing independently selected from —F, —Cl, —Br, —I, —OH, —NO₂, —CN, or—NH₂.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

It is understood that any alkyl, alkenyl, alkynyl, cyclically andcycloalkenyl moiety described herein can also be an aliphatic group, analicyclic group or a heterocyclic group. An “aliphatic group” isnon-aromatic moiety that may contain any combination of carbon atoms,hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, andoptionally contain one or more units of unsaturation, e.g., doubleand/or triple bonds. An aliphatic group may be straight chained,branched or cyclic and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, aliphatic groups include,for example, polyalkoxyalkyls, such as polyalkylene glycols, polyamines,and polyimines, for example. Such aliphatic groups may be furthersubstituted. It is understood that aliphatic groups may be used in placeof the alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylenegroups described herein.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or polycyclic saturated carbocyclic ring compound bythe removal of a single hydrogen atom. Examples include, but not limitedto, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [2.2.1]heptyl, and bicyclo [2.2.2] octyl. Such alicyclic groups may be furthersubstituted.

The term “heterocycloalkyl” and “heterocyclic” can be usedinterchangeably and refer to a non-aromatic 3-, 4-, 5-, 6- or 7-memberedring or a bi- or tri-cyclic group fused system, where (i) each ringcontains between one and three heteroatoms independently selected fromoxygen, sulfur and nitrogen, (ii) each 5-membered ring has 0 to 1 doublebonds and each 6-membered ring has 0 to 2 double bonds, (iii) thenitrogen and sulfur heteroatoms may optionally be oxidized, (iv) thenitrogen heteroatom may optionally be quaternized, (v) any of the aboverings may be fused to a benzene ring, and (vi) the remaining ring atomsare carbon atoms which may be optionally oxo-substituted. Representativeheterocycloalkyl groups include, but are not limited to, [1,3]dioxolane,pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl,thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, andtetrahydrofuryl. Such heterocyclic groups may be further substituted togive substituted heterocyclic.

It will be apparent that in various embodiments of the invention, thesubstituted or unsubstituted alkyl, alkenyl, alkynyl, cyclically,cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl, andheterocycloalkyl are intended to be monovalent or divalent. Thus,alkylene, alkenylene, and alkynylene, cycloaklylene, cycloalkenylene,cycloalkynylene, arylalkylene, hetoerarylalkylene andheterocycloalkylene groups are to be included in the above definitions,and are applicable to provide the formulas herein with proper valency.

The term “hydroxy activating group”, as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxy group sothat it will depart during synthetic procedures such as in asubstitution or elimination reactions. Examples of hydroxy activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy groupactivated with a hydroxy activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups.

The terms “halo” and “halogen,” as used herein, refer to an atomselected from fluorine, chlorine, bromine and iodine.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques, which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included. Theconfiguration of any carbon-carbon double bond appearing herein isselected for convenience only and is not intended to designate aparticular configuration unless the text so states; thus a carbon-carbondouble bond depicted arbitrarily herein as trans may be cis, trans, or amixture of the two in any proportion.

The term “subject” as used herein refers to a mammal. A subjecttherefore refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, and the like. Preferably the subject is a human. When the subjectis a human, the subject may be referred to herein as a patient.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxy groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theare described generally in T. H. Greene and P. G., S. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxy protecting groups includebenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl,2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl,trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl,2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, 1,1-dimethyl-2-propenyl,3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl,triphenylmethyl (trityl), tetrahydrofuryl, methoxymethyl,methylthiomethyl, benzyloxymethyl, 2,2,2-triehloroethoxymethyl,2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl,trimethylsilyl, triethylsilyl, triisopropylsilyl, and the like.Preferred hydroxy protecting groups for the present invention are acetyl(Ac or —C(O)CH₃), benzoyl (Bz or —C(O)C₆H₅), and trimethylsilyl (TMS or—Si(CH₃)₃). Berge, et al. describes pharmaceutically acceptable salts indetail in J. Pharmaceutical Sciences, 66: 1-19 (1977). The salts can beprepared in situ during the final isolation and purification of thecompounds of the invention, or separately by reacting the free basefunction with a suitable organic acid. Examples of pharmaceuticallyacceptable salts include, but are not limited to, nontoxic acid additionsalts e.g., salts of an amino group formed with inorganic acids such ashydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid andperchloric acid or with organic acids such as acetic acid, maleic acid,tartaric acid, citric acid, succinic acid or malonic acid or by usingother methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include, but are not limited to,adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the aredescribed generally in T. H. Greene and P. G. M. Wuts, Protective Groupsin Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999).Examples of amino protecting groups include, but are not limited to,t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and thelike.

As used herein, the term “pharmaceutically acceptable ester” refers toesters of the compounds formed by the process of the present inventionwhich hydrolyze in vivo and include those that break down readily in thehuman body to leave the parent compound or a salt thereof. Suitableester groups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Examples ofparticular esters include, but are not limited to, formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the present invention. “Prodrug”, as used hereinmeans a compound, which is convertible in vivo by metabolic means (e.g.by hydrolysis) to afford any compound delineated by the formulae of theinstant invention. Various forms of prodrugs are known in the art, forexample, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier(1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4, AcademicPress (1985); Krogsgaard-Larsen, et al., (ed). “Design and Applicationof Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et al., Journal of Drug Deliver Reviews,8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq.(1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug DeliverySystems, American Chemical Society (1975); and Bernard Testa & JoachimMayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

The term “acyl” includes residues derived from acids, including but notlimited to carboxylic acids, carbamic acids, carbonic acids, sulfonicacids, and phosphorous acids. Examples include aliphatic carbonyls,aromatic carbonyls, aliphatic sulfonyls, aromatic sulfinyls, aliphaticsulfinyls, aromatic phosphates and aliphatic phosphates. Examples ofaliphatic carbonyls include, but are not limited to, acetyl, propionyl,2-fluoroacetyl, butyryl, 2-hydroxy acetyl, and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofaprotic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

The terms “protogenic organic solvent” or “protic solvent” as usedherein, refer to a solvent that tends to provide protons, such as analcohol, for example, methanol, ethanol, propanol, isopropanol, butanol,t-butanol, and the like. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofprotogenic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. Additionally, thevarious synthetic steps may be performed in an alternate sequence ororder to give the desired compounds. In addition, the solvents,temperatures, reaction durations, etc. delineated herein are forpurposes of illustration only and variation of the reaction conditionscan produce the desired bridged macrocyclic products of the presentinvention. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein include, for example, those described in R.Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d.Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser andFieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); andL. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, JohnWiley and Sons (1995).

The compounds of this invention may be modified by appending variousfunctionalities via synthetic means delineated herein to enhanceselective biological properties. Such modifications include those whichincrease biological penetration into a given biological system (e.g.,blood, lymphatic system, central nervous system), increase oralavailability, increase solubility to allow administration by injection,alter metabolism and alter rate of excretion.

PHARMACEUTICAL COMPOSITIONS

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

ANTIVIRAL ACTIVITY

An inhibitory amount or dose of the compounds of the present inventionmay range from about 0.01 mg/Kg to about 500 mg/Kg, alternatively fromabout 1 to about 50 mg/Kg. Inhibitory amounts or doses will also varydepending on route of administration, as well as the possibility ofco-usage with other agents.

According to the methods of treatment of the present invention, viralinfections are treated or prevented in a subject such as a human orlower mammal by administering to the subject an anti-hepatitis C virallyeffective amount or an inhibitory amount of a compound of the presentinvention, in such amounts and for such time as is necessary to achievethe desired result. An additional method of the present invention is thetreatment of biological samples with an inhibitory amount of a compoundof composition of the present invention in such amounts and for suchtime as is necessary to achieve the desired result.

The term “anti-hepatitis C virally effective amount” of a compound ofthe invention, as used herein, mean a sufficient amount of the compoundso as to decrease the viral load in a biological sample or in a subject(e.g., resulting in at least 10%, preferably at least 50%, morepreferably at least 80%, and most preferably at least 90% or 95%,reduction in viral load). As well understood in the medical arts, ananti-hepatitis C virally effective amount of a compound of thisinvention will be at a reasonable benefit/risk ratio applicable to anymedical treatment.

The term “inhibitory amount” of a compound of the present inventionmeans a sufficient amount to decrease the hepatitis C viral load in abiological sample or a subject (e.g., resulting in at least 10%,preferably at least 50%, more preferably at least 80%, and mostpreferably at least 90% or 95%, reduction in viral load). It isunderstood that when said inhibitory amount of a compound of the presentinvention is administered to a subject it will be at a reasonablebenefit/risk ratio applicable to any medical treatment as determined bya physician. The term “biological sample(s),” as used herein, means asubstance of biological origin intended for administration to a subject.Examples of biological samples include, but are not limited to, bloodand components thereof such as plasma, platelets, subpopulations ofblood cells and the like; organs such as kidney, liver, heart, lung, andthe like; sperm and ova; bone marrow and components thereof, or stemcells. Thus, another embodiment of the present invention is a method oftreating a biological sample by contacting said biological sample withan inhibitory amount of a compound or pharmaceutical composition of thepresent invention.

Upon improvement of a subject's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease. Thesubject may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

It will be understood, however, that the total daily usage of thecompounds and compositions of the present invention will be decided bythe attending physician within the scope of sound medical judgment. Thespecific inhibitory dose for any particular patient will depend upon avariety of factors including the disorder being treated and the severityof the disorder; the activity of the specific compound employed; thespecific composition employed; the age, body weight, general health, sexand diet of the patient; the time of administration, route ofadministration, and rate of excretion of the specific compound employed;the duration of the treatment; drugs used in combination or coincidentalwith the specific compound employed; and like factors well known in themedical arts.

The total daily inhibitory dose of the compounds of this inventionadministered to a subject in single or in divided doses can be inamounts, for example, from 0.01 to 50 mg/kg body weight or more usuallyfrom 0.1 to 25 mg/kg body weight. Single dose compositions may containsuch amounts or submultiples thereof to make up the daily dose. Ingeneral, treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one with ordinary skill inthe art. All publications, patents, published patent applications, andother references mentioned herein are hereby incorporated by referencein their entirety.

ABBREVIATIONS

Abbreviations which have been used in the descriptions of the schemesand the examples that follow are:

-   -   ACN for acetonitrile;    -   BME for 2-mercaptoethanol;    -   BOP for benzotriazol-1-yloxy-tris(dimethylamino)phosphonium        hexafluorophosphate;    -   COD for cyclooctadiene;    -   DAST for diethylaminosulfur trifluoride;    -   DABCYL for        6-(N-4′-carboxy-4-(dimethylamino)azobenzene)-aminohexyl-1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite;    -   DCM for dichloromethane;    -   DIAD for diisopropyl azodicarboxylate;    -   DIBAL-H for diisobutylaluminum hydride;    -   DIEA for diisopropyl ethylamine;    -   DMAP for N,N-dimethylaminopyridine;    -   DME for ethylene glycol dimethyl ether;    -   DMEM for Dulbecco's Modified Eagles Media;    -   DMF for N,N-dimethyl formamide;    -   DMSO for dimethylsulfoxide;    -   DUPHOS for

-   -   EDANS for 5-(2-Amino-ethylamino)-naphthalene-1-sulfonic acid;    -   EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide        hydrochloride;    -   EtOAc for ethyl acetate;    -   HATU for O(7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluorophosphate;    -   Hoveyda's Cat. for Dichloro(o-isopropoxyphenylmethylene)        (tricyclohexylphosphine)ruthenium(II);    -   KHMDS is potassium bis(trimethylsilyl) amide;    -   Ms for mesyl;    -   NMM for N-4-methylmorpholine;    -   PyBrOP for Bromo-tri-pyrolidino-phosphonium hexafluorophosphate;    -   Ph for phenyl;    -   RCM for ring-closing metathesis;    -   RT for reverse transcription;    -   RT-PCR for reverse transcription-polymerase chain reaction;    -   TEA for triethyl amine;    -   TFA for trifluoroacetic acid;    -   THF for tetrahydrofuran;    -   TLC for thin layer chromatography;    -   TPP or PPh₃ for triphenylphosphine;    -   tBOC or Boc for tert-butyloxy carbonyl; and    -   Xantphos for        4,5-Bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene.

SYNTHETIC METHODS

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared, which are intended as an illustration only and not to limitthe scope of the invention. Various changes and modifications to thedisclosed embodiments will be apparent to those skilled in the art andsuch changes and modifications including, without limitation, thoserelating to the chemical structures, substituents, derivatives, and/ormethods of the invention may be made without departing from the spiritof the invention and the scope of the appended claims.

The preparation of triazole-containing macrocyclic compounds isexemplified in Scheme 1. Boc-hydroxyproline mesylate 1-1 reacted withtriazole derivative 1-2 (for the preparation, see Scheme 3) under SN₂basic conditions giving triazole compound 1-3. Introduction of vinylgroup is accomplished via Suzuki (vinylborate/palladium) or Still(vinyltin/Palladium) reaction to afford compound 1-4. Deprotection of1-4 with HCl followed by coupling reaction (HATU/DMF) with compound 1-5(for the preparation of carbamate aminoacid derivatives see Scheme 6)resulted in the macrocyclic compound precursor 1-6. Ring-closingmetathesis of compound 1-6 with a ruthenium-based catalyst gave thedesired macrocyclic intermediate 1-7 (for further details on ringclosing metathesis see recent reviews: Grubbs et al., Acc. Chem. Res.,1995, 28, 446; Shrock et al., Tetrahedron 1999, 55, 8141; Furstner, A.Angew. Chem. Int. Ed. 2000, 39, 3012; Trnka et al., Acc. Chem. Res.2001, 34, 18, and Hoveyda et al., Chem. Eur. J. 2001, 7, 945).

The syntheses of triazole-containing macrocyclic compounds as HCVprotease inhibitors are exemplified in Scheme 2. Hydrolysis of compound1-7 gives the corresponding carboxylic acid 1-8, which is coupled withamino sulfinimide 1-9 or amino acid ester 1-10 to afford compound 1(example 1) or ester 1-11. Hydrolysis of 1-11 gives 1-12. Compound 1 canalso be prepared from the acid 1-12 as shown in scheme 2. Hydrogenationof 1-7 gives compound 1-13, which is is coupled with amino sulfinimide1-9 to afford compounds 1-14 (example 2). Coupling of compound 1-8 andcompound 1-15 results in compounds 1-16.

Triazoles of the present invention are prepared by reacting alkynecompound, which is commercial available or made according to Scheme 4,and trimethylsilyl azide via the general synthetic route described inScheme 3 and 4. For further details concerning 1,2,3 Triazoles fromsubstituted alkynes and TMS-N₃ see Bickofer et al., Chem. Ber. 1966, 99,2512-2517.

Synthesis of Alkynes Via Sonogashira Reaction

-   -   Alkynes used in the present invention can be made by the        Sonogashira reaction with primary alkyne compound, aryl halide        (Y-halide), base, palladium catalyst and CuI via the synthetic        route described generally in Scheme 4. For further details of        the Sonogashira reaction see Sonogashira, Comprehensive Organic        Synthesis, Volume 3, Chapters 2,4 and Sonogashira, Synthesis        1977, 777.

Synthesis of Alkynyl Amides

Additional alkynes used in the present invention can be made by reactingalkynyl acid with amine in the presence of a peptide coupling reagent,such as BOP, HATU, PryBOP, and the like, and a base, such as DIEA, inDMF via the synthetic route described generally in Scheme 5.

Synthesis of Carbamate Amino Acids

The preparation of carbamate amino acids (compound 1-5) is shown inScheme 6. The corresponding olefine alcohol reactes with triphosphogenin presence of DIPEA, followed by adding the appropriate amino acids togive the desired carbamate aminoacids.

All references cited herein, whether in print, electronic, computerreadable storage media or other form, are expressly incorporated byreference in their entirety, including but not limited to, abstracts,articles, journals, publications, texts, treatises, internet web sites,databases, patents, and patent publications.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not to limit the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Example 1

Compound of Formula VII, wherein

Step 1A

-   -   To a mixture of 1-bromo-3-iodobenzene (3.8 ml, 30 mmol),        phenylacetylene (1.65 ml, 15 mmol), copper iodide (285 mg, 1.5        mmol), THF (20 ml) and TMEDA (20 m) was added        tetrakis(triphenylphosphine)palladium (0) (620 mg, 0.54 mmol).        The resulting mixture was stirred at 50° C. for 20 h, cooled to        room temperature, diluted with EtOAc (250 ml), washed with aq.        NaHCO3, brine, dried (MgSO4), concentrated under vacuum and the        residue was purified by chromatography (Hexane) to give 1a (2.7        g).

Step 1B

-   -   A mixture of compound 1a (0.5 g, 1.94 mmol), trimethylsilyl        azide (0.9 ml, 6.8 mmol) and DMF (3 ml) was stirred at 135° C.        for 36 h, cooled to room temperature, diluted with EtOAc, washed        brine (3×), dried (MgSO4), concentrated under vacuum and the        residue was purified by chromatography (Hexane/AcOEt=4:1 to 3:1)        to give 1b (350 mg). MS(ESI): m/z 300.06, 302.03 (M+H).

Step 1C

-   -   A mixture of compound 1b (339 mgg, 1.13 mmol), compound 1c-1        (401 mg, 1.24 mmol), cesium carbonate (736 mg, 2.26 mmol) and        DMF (5 ml) was stirred at 70° C. for 20 h, diluted with EtOAc,        washed with brine (3×), dried (MgSO4) and concentrated in vacuo.        The residue was purified by silica gel chromatography        (Hexane/EtOAc=9:1 to 8:2) to afford 1c (373 mg). MS(ESI): m/z        527.20, 529.20 (M+H).

Step 1D

To a mixture of compound 1c (365 mgg, 0.69 mmol), potassiumvinyltrifluoroborate (280 mg, 2.1 mmol), triethylamine (0.3 ml) andethanol (8 ml) was added 1,1′-bis(diphenylphopshino)ferrocene palladium(II) chloride complex with CH₂Cl₂(50 mg, 0.055 mmol). The resultingmixture was stirred at 75° C. for 15 h, cooled to room temperature,quenched with 10% KHSO4 aq. solution, extracted with ethyl acetate (3×).The combined oranic layers were dried (MgSO4), concentrated undervacuum, and the residue was purified by chromatography (Hexane/EtOAC=1:0to 4:1) to give 1d (226 g). MS(ESI): m/z 475.14 (M+H), 513.08 (M+K).

Step 1E

A solution of compound 1d (220 mg, 0.47 mmol) in dichloromethane (1 ml)was treated with 4M HCl/dioxane (3 ml, 12 mmol). The resulting mixturewas stirred at room temperature for 1 hour, and concentrated in vacuo todryness to afford HCl salt of compound 1e (100%). MS (ESI): m/e 375.07(M+H).

Step 1F

To a solution of hex-5-en-1-ol (1.3 ml, 10.9 mol), triphosphogen (1.46g, 4.92 mmol) in 1,4-dioxane (21 ml) at 0° C. was added dropwise DIPEA(1.7 ml, 9.72 mmol). The mixture was stirred at room temperature for anhour, cooled to 0° C. To this was added slowly L-t-leucine (1.28 g, 9.72mmol) dissolved in 1N NaOH (9.8 ml). The resulting mixture was stirredat room temperature overnight, concentrated to remove half volume ofdioxane, treated with 1N NaOH (25 ml), washed with ether (3×30 ml). Theaqueous phase was acidified to pH 2˜3 with 6N HCl, then extracted withdichloromethane (3×30 ml). The combined oranic layers were dried(MgSO4), concentrated to dryness to give compound 1f (2.5 g) directlyused in next step.

Step 1G

To a solution of compound 1e (HCl salt, 0.23 mmol), compound 1f (90 mg,0.35 mmol) and DIPEA (0.16 ml, 0.92 mmol) in DMF (3 ml) at 0° C. wasadded HATU (140 mg, 0.37 mmol). The mixture was stirred at rt for 18 h,diluted with EtOAc and washed with half-sat.-aq. NaCl four times. Theorganic phase was dried over anhydrous MgSO₄, filtered, and thenconcentrated in vacuo. The resudue was purified by silica gelchromatography (Hexane/EtOAC=9:1 to 7:3) to afford compound 1g(130 mg).MS (ESI): m/z 614.24 (M+H).

Step 1H

To a solution of compound 1g (47 mg, 0.19 mmol) in dichloromethane (20ml) was added Hoveyda-Grubbs' 1^(st) generation catalyst (5 mol % eq.).The reaction mixture was stirred at 40° C. for 20 h. The solvent wasthen evaporated and the residue was purified by silica gel flashchromatography using gradient elution (Hexane/EtOAC=9:1 to 7:3) to yieldthe macrocyclic compound 1f (25 mg). MS (ESI) m/z 586.27 (M+H).

Step 1I

-   -   To a solution of compound 1h (20 mg, 0.034 mmol) in THF/MeOH (4        ml/2 ml) was added 1N lithium hydroxide (2 ml, 2 mmol). The        mixture was stirred at room temperature for 20 hours. Most        organic solvents were evaporated in vacuo, and the resulting        residue was diluted with water and acidified to pH 5 to 6. The        mixture was extracted with EtOAc three times. The combined        organic extracts were dried (MgSO₄), filtered and concentrated        in vacuo to afford 1i (100%). MS(ESI): m/z 572.28 (M+H).

Step 1J

To a solution of compound 1i (0.035 mmol), compound 1j-1 (0.08 M in DMF,0.77 ml, 0.06 mmol) and DIPEA (0.065 ml, 0.373 mmol) in DMF (3 ml) at 0°C. was added HATU (30 mg, 0.078 mmol). The mixture was stirred at rt for18 h, diluted with EtOAc and washed with half-sat.-aq. NaCl four times.The organic phase was dried over anhydrous MgSO₄, filtered, and thenconcentrated in vacuo. The resudue was purified by preparative HPLC toafford the title compound (11 mg). MS (ESI): m/z 784.33 (M+H).

Example 2

Compound of Formula VII, wherein

Step2A

A mixture of compound 1i (13 mg, 0.022 mmol), Pd—C (10%, 4 mg) andmethanol (4 ml) was hydrogenated under atmopheric pressure for 1 h,filtered, washed with ethyl acetate. The filtrate was concentrated todryness to give compound 2a used directy in next step. MS (ESI): m/z574.26 (M+H).

Step2B

-   -   To a solution of the avove compound 2a (10 mg, 0.017 mmol),        compound 1j-1 (0.08 M in DMF, 0.4 ml, 0.03 mmol) and DIPEA        (0.055 ml, 0.315 mmol) in DMF (1ml) at 0° C. was added HATU (16        mg, 0.043 mmol). The mixture was stirred at rt for 18 h, diluted        with EtOAc and washed with half-sat.-aq. NaCl four times. The        organic phase was dried over anhydrous MgSO₄, filtered, and then        concentrated in vacuo. The resudue was purified by preparative        HPLC to afford the title compound (4 mg). MS (ESI): m/z 786.34        (M+H).

Example 3

Compound of Formula VII, wherein

A mixture of compound example 1 (5 mg, 0.006 mmol), Pd—C (10%, 4 mg) andethyl acetate (4 ml) was hydrogenated under atmopheric pressure for 2 h,filtered, washed with ethyl acetate. The filtrate was concentrated andthe resudue was purified by preparative HPLC to afford the titlecompound (3 mg). MS (ESI): m/z 788.34 (M+H).

Example 4 to 144

Compounds of Formula VII in Table 1, are made following the proceduresdescribed in Example 1 to 3 and the Synthetic Methods section.

The compounds of the present invention exhibit potent inhibitoryproperties against the HCV NS3 protease and ahowed activities inreplicon assays. The following examples describe assays in which thecompounds of the present invention can be tested for anti-HCV effects.

Example 145 NS3/NS4a Protease Enzyme Assay

HCV protease activity and inhibition is assayed using an internallyquenched fluorogenic substrate. A DABCYL and an EDANS group are attachedto opposite ends of a short peptide. Quenching of the EDANS fluorescenceby the DABCYL group is relieved upon proteolytic cleavage. Fluorescenceis measured with a Molecular Devices Fluoromax (or equivalent) using anexcitation wavelength of 355 nm and an emission wavelength of 485 nm.

The assay is run in Corning white half-area 96-well plates (VWR29444-312 [Corning 3693]) with full-length NS3 HCV protease 1b tetheredwith NS4A cofactor (final enzyme concentration 1 to 15 nM). The assaybuffer is complemented with 10 μM NS4A cofactor Pep 4A (Anaspec 25336 orin-house, MW 1424.8). RET S1 (Ac-Asp-Glu-Asp(EDANS)-Glu-Glu-Abu-[COO]Ala-Ser-Lys-(DABCYL)-NH₂ (SEQ ID NO: 4), AnaSpec22991, MW 1548.6) is used as the fluorogenic peptide substrate. Theassay buffer contains 50 mM Hepes at pH 7.5, 30 mM NaCl and 10 mM BME.The enzyme reaction is followed over a 30 minutes time course at roomtemperature in the absence and presence of inhibitors.

The peptide inhibitors HCV Inh 1 (Anaspec 25345, MW 796.8)Ac-Asp-Glu-Met-Glu-Glu-Cys-OH (SEQ ID NO: 5), [−20° C.] and HCV Inh 2(Anaspec 25346, MW 913.1) Ac-Asp-Glu-Dif-Cha-Cys-OH (SEQ ID NO: 6), areused as reference compounds.

IC50 values are calculated using XLFit in ActivityBase (IDBS) usingequation 205:y=A+((B−A)/(1+((C/x)^D))).

Example 146 Cell-Based Replicon Assay

Quantification of HCV replicon RNA (HCV Cell Based Assay) isaccomplished using the Huh 11-7 cell line (Lohmann, et al Science285:110-113, 1999). Cells are seeded at 4×10³ cells/well in 96 wellplates and fed media containing DMEM (high glucose), 10% fetal calfserum, penicillin-streptomycin and non-essential amino acids. Cells areincubated in a 7.5% CO₂ incubator at 37° C. At the end of the incubationperiod, total RNA is extracted and purified from cells using AmbionRNAqueous 96 Kit (Catalog No. AM 1812). To amplify the HCV RNA so thatsufficient material can be detected by an HCV specific probe (below),primers specific for HCV (below) mediate both the reverse transcriptionof the HCV RNA and the amplification of the cDNA by polymerase chainreaction (PCR) using the TaqMan One-Step RT-PCR Master Mix Kit (AppliedBiosystems catalog no. 4309169). The nucleotide sequences of the RT-PCRprimers, which are located in the NS5B region of the HCV genome, are thefollowing:

HCV Forward primer “RBNS5bfor” 5′GCTGCGGCCTGTCGAGCT: (SEQ ID NO: 1) HCVReverse primer “RBNS5Brev” 5′CAAGGTCGTCTCCGCATAC. (SEQ ID NO 2)

Detection of the RT-PCR product is accomplished using the AppliedBiosystems (ABI) Prism 7500 Sequence Detection System (SDS) that detectsthe fluorescence that is emitted when the probe, which is labeled with afluorescence reporter dye and a quencher dye, is degraded during the PCRreaction. The increase in the amount of fluorescence is measured duringeach cycle of PCR and reflects the increasing amount of RT-PCR product.Specifically, quantification is based on the threshold cycle, where theamplification plot crosses a defined fluorescence threshold. Comparisonof the threshold cycles of the sample with a known standard provides ahighly sensitive measure of relative template concentration in differentsamples (ABI User Bulletin #2 Dec. 11, 1997). The data is analyzed usingthe ABI SDS program version 1.7. The relative template concentration canbe converted to RNA copy numbers by employing a standard curve of HCVRNA standards with known copy number (ABI User Bulletin #2 Dec. 11,1997).

The RT-PCR product was detected using the following labeled probe:

(SEQ ID NO: 3) 5′FAM-CGAAGCTCCAGGACTGCACGATGCT-TAMRA FAM = Fluorescencereporter dye. TAMRA: = Quencher dye.

The RT reaction is performed at 48° C. for 30 minutes followed by PCR.Thermal cycler parameters used for the PCR reaction on the ABI Prism7500 Sequence Detection System are: one cycle at 95° C., 10 minutesfollowed by 40 cycles each of which include one incubation at 95° C. for15 seconds and a second incubation for 60° C. for 1 minute.

To normalize the data to an internal control molecule within thecellular RNA, RT-PCR is performed on the cellular messenger RNAglyceraldehyde-3-phosphate dehydrogenase (GAPDH). The GAPDH copy numberis very stable in the cell lines used. GAPDH RT-PCR is performed on thesame RNA sample from which the HCV copy number is determined. The GAPDHprimers and probesare contained in the ABI Pre-Developed TaqMan AssayKit (catalog no. 4310884E). The ratio of HCV/GAPDH RNA is used tocalculate the activity of compounds evaluated for inhibition of HCV RNAreplication.

Activity of Compounds as Inhibitors of HCV Replication (Cell BasedAssay) in Replicon Containing Huh-7 Cell Lines

The effect of a specific anti-viral compound on HCV replicon RNA levelsin Huh-11-7 cells is determined by comparing the amount of HCV RNAnormalized to GAPDH (e.g. the ratio of HCV/GAPDH) in the cells exposedto compound versus cells exposed to the DMSO vehicle (negative control).Specifically, cells are seeded at 4×10³ cells/well in a 96 well plateand are incubated either with: 1) media containing 1% DMSO (0%inhibition control), or 2) media/1% DMSO containing a fixedconcentration of compound. 96 well plates as described above are thenincubated at 37° C. for 4 days (EC50 determination). Percent inhibitionis defined as:% Inhibition=100−100*S/C1

-   -   where    -   S=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        sample;    -   C1=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        0% inhibition control (media/1% DMSO).        The dose-response curve of the inhibitor is generated by adding        compound in serial, three-fold dilutions over three logs to        wells starting with the highest concentration of a specific        compound at 1.5 uM and ending with the lowest concentration of        0.23 nM. Further dilution series (500 nM to 0.08 nM for example)        is performed if the EC50 value is not positioned well on the        curve. EC50 is determined with the IDBS Activity Base program        “XL Fit” using a 4-paramater, non-linear regression fit (model #        205 in version 4.2.1, build 16).

1. A compound of Formula I, II or III:

or a pharmaceutically acceptable salt thereof, wherein A is absent orselected from (C═O), S(O)₂, C(═N—OR₁) or C(═N—CN); wherein R₁ isselected at each occurrence from the group consisting of: (i) hydrogen;(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl; (iii)heterocycloalkyl or substituted heterocycloalkyl; (iv) —C₁-C₈ alkyl,—C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl each eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; L₂₀₁ is absent or selected from C₁-C₈alkylene, C₂-C₈ alkynylene containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; substituted C₁-C₈ alkylene, substituted C₂-C₈alkenylene, or substituted C₂-C₈ alkynylene each containing 0, 1, 2, or3 heteroatoms selected from O, S or N; C₃C₁₂ cycloalkylene, orsubstituted C₃-C₁₂ cycloalkylene each containing 0, 1, 2, or 3heteroatoms selected from O, S or N; C₃-C₁₂ cycloalkenylene, orsubstituted C₃C₁₂ cycloalkenylene each containing 0, 1, 2, or 3heteroatoms selected from O, S or N; M is absent or selected from O, Sor NR₁; L₁₀₁ is absent or selected from C₁-C₈ alkylene, C₂-C₈alkenylene, or C₂-C₈ alkynylene each containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; substituted C₁-C₈ alkylene,substituted C₂-C₈ alkenylene, or substituted C₂-C₈ alkynylene eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; C₃C₁₂cycloalkylene, or substituted C₃-C₁₂ cycloalkylene each containing 0, 1,2, or 3 heteroatoms selected from O, S or N; C₃-C₁₂ cycloalkenylene, orsubstituted C₃C₁₂ cycloalkenylene each containing 0, 1, 2, or 3heteroatoms selected from O, S or N; Z₁₀₁ is absent or selected fromaryl, substituted aryl, heteroaryl, or substituted heteroaryl; W₁₀₁ isabsent, or selected from C₁-C₈ alkylene, C₂-C₈ alkenylene, or C₂-C₈alkynylene, C(O)NR₁, C(O), aryl, substituted aryl, heteroaryl, orsubstituted heteroaryl; W₂₀₁ is selected from hydrogen, halogen, C₁-C₆alkyl, C₃-C₁₂ cycloalkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl,substituted heteroarylalkyl, heterocycloalkyl, or substitutedheterocycloalkyl; alternatively, W₁₀₁ and W₂₀₁ taken together with thecarbon atoms to which they are attached to form a cyclic moiety selectedfrom aryl, substituted aryl, heteroaryl, or substituted heteroaryl; Rand R′ are independently selected from the group consisting of: (i)—C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2,or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₄-C₁₂ alkylcycloalkyl,or substituted —C₄-C₁₂ alkylcycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; —C₄-C₁₂ alkylcycloalkenyl, orsubstituted —C₄-C₁₂ alkylcycloalkenyl; (ii) aryl; substituted aryl;heteroaryl; substituted heteroaryl; (iii) heterocycloalkyl orsubstituted heterocycloalkyl; (iv) hydrogen; deuterium; G is selectedfrom —OH, —NR₃R₄, —NHS(O)₂—R₂, —NH(SO₂)NR₃R₄; R₂ is selected from: (i)aryl; substituted aryl; heteroaryl; substituted heteroaryl; (ii)heterocycloalkyl or substituted heterocycloalkyl; (iii) —C₁-C₈ alkyl,—C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S or N, substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃C₁₂ cycloalkenyl; R₃ and R₄ are independently selectedfrom: (i) hydrogen; (ii) aryl; substituted aryl; heteroaryl; substitutedheteroaryl; (iii) heterocycloalkyl or substituted heterocycloalkyl; (iv)—C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2,or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl each containing0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl,or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or substituted—C₃-C₁₂ cycloalkenyl; m =0, 1, or 2; and m′=1 or
 2. 2. The compound ofclaim 1, wherein the compound is of Formula IV, V or VI:

or a pharmaceutically acceptable salt thereof, alone or in combinationwith a pharmaceutically acceptable carrier or excipient, where R, R′, A,L₂₀₁, M, L₁₀₁, Z₁₀₁, W₁₀₁, W₂₀₁ and G are as previously defined inclaim
 1. 3. A compound according to claim 1 which is selected fromcompounds of Formula VII and pharmaceutically acceptable salts thereofwherein R, L-W, Z, R′ and G are delineated in Table 1: TABLE 1 (VII)

Example # R L-W Z R′ G
 1.


2.


3.


4.

OH
 5.


6.


7.


8.

OH
 9.


10.


11.


12.

OH
 13.


14.


15.


16.

OH
 17.


18.


19.


20.

OH
 21.


22.


23.


24.

OH
 25.


26.


27.


28.

OH
 29.


30.


31.


32.

OH
 33.


34.


35.


36.

OH
 37.


38.


39.


40.

OH
 41.


42.


43.


44.

OH
 45.


46.


47.


48.

OH
 49.


50.


51.


52.

OH
 53.


54.


55.


56.

OH
 57.


58.


59.


60.

OH
 61.


62.


63.


64.

OH
 65.


66.


67.


68.

OH
 69.


70.


71.


72.

OH
 73.


74.


75.


76.

OH
 77.


78.


79.


80.

OH
 81.


82.


83.


84.

OH
 85.


86.


87.


88.

OH
 89.


90.


91.


92.

OH
 93.


94.


95.


96.

OH
 97.


98.


99.


100.

OH
 101.


102.


103.


104.

OH
 105.


106.


107.


108.

OH
 109.


110.


111.


112.

OH
 113.


114.


115.


116.

OH
 117.


118.


119.


120.

OH
 121.


122.


123.


124.

OH
 125.


126.


127.


128.

OH
 129.


130.


131.


132.

OH
 133.


134.


135.


136.

OH
 137.


138.


139.


140.

OH
 141.


142.


143.


144.

OH.


4. A pharmaceutical composition comprising an inhibitory amount of acompound according to claim 1 or a pharmaceutically acceptable saltthereof, in combination with a pharmaceutically acceptable carrier orexcipient.
 5. A method of treating a viral infection in a subject,comprising administering to the subject an inhibitory amount of apharmaceutical composition according to claim
 4. 6. The method accordingto claim 5, wherein the viral infection is hepatitis C virus.
 7. Amethod of inhibiting the replication of hepatitis C virus, the methodcomprising supplying a hepatitis C viral NS3 protease inhibitory amountof the pharmaceutical composition of claim
 5. 8. The method of claim 5further comprising administering concurrently an additionalanti-hepatitis C virus agent.
 9. The method of claim 8, wherein saidadditional anti-hepatitis C virus agent is selected from the groupconsisting of α-interferon, β-interferon, ribavarin, and adamantine. 10.The method of claim 8, wherein said additional anti-hepatitis C virusagent is an inhibitor of hepatitis C virus helicase, polymerase,metalloprotease, or IRES.
 11. The pharmaceutical composition of claim 4,further comprising another anti-HCV agent.
 12. The pharmaceuticalcomposition of claim 4, further comprising an agent selected frominterferon, ribavirin, amantadine, another HCV protease inhibitor, anHCV polymerase inhibitor, an HCV helicase inhibitor, or an internalribosome entry site inhibitor.
 13. The pharmaceutical composition ofclaim 4, further comprising pegylated interferon.
 14. The pharmaceuticalcomposition of claim 4, further comprising another anti-viral,anti-bacterial, anti-fungal or anti-cancer agent, or an immunemodulator.